Infrared signal monitoring for cell cultures

ABSTRACT

The present invention is directed to an apparatus for monitoring a cell culture comprising a) one or more infrared sensors positioned adjacent to a cell culture, the one or more infrared sensors capable of recording an infrared heat signal from the cell culture; b) a power source in electrical communication with the one or more infrared sensors; c) a data storage and computation device configured to receive and analyze the infrared heat signal from the one or more infrared sensors; and d) a transmitter device in electrical communication with the storage and computation device; wherein the apparatus monitors the pattern of heat production in the cell culture. The present invention is also directed to methods for monitoring and analyzing the metabolic activity of cells using the above apparatus.

This application is a continuation of U.S. application Ser. No.14/965,321, titled “INFRARED SIGNAL MONITORING FOR CELL CULTURES” filedDec. 10, 2015, the contents of which are incorporated by referenceherein in its entirety.

BACKGROUND

The present invention relates to a system for monitoring of cellcultures, and in particular to a system of monitoring cell culturesusing infrared sensors and cameras to detect changes in the pattern ofheat production within metabolically active cells.

Cells are commonly grown in the lab on dishes or in wells that containnutrients. These may be human or animal cells, as well as microorganismssuch as bacteria, fungi and archaea used in research or clinicaldiagnostics and therapeutics. Cells are typically grown in an incubatorwith controlled temperature and humidity. To determine the degree ofgrowth of cells, they are most often monitored visually by a human. Forexample, samples from potential infection sites in patients areinoculated on a Petri dish and a lab technician may visually inspect thedish after a few hours to a few days to spot signs of bacterial colonyformation. This is labor intensive and also is associated with a riskfor contamination. When cells require a strict anaerobic environment,visually inspecting them entails exposing them (even if for a limitedperiod of time) to air, which may delay their growth or lead to theirdeath. Visual inspection is widely used to determine bacterialsusceptibility to antibiotics, for example using the disc-diffusiontest, wherein bacterial growth is inhibited around a disc containing anantibiotic drug. However, it is practically impossible for humans tocontinuously monitor cells in culture. Moreover, for microorganismgrowth to be visually evident to the naked eye takes an extremely highnumber of cells to be accumulated.

Yet the ability to continuously monitor cells in culture is desirable.For example, it could shorten the time until an infection is diagnosedor the time needed to determine what the most effective antibiotic wouldbe to treat a patient. Moreover, cells may stop growing or die due tolack of nutrients, unbalanced ambient conditions or infection byviruses, phages and fungi. Early detection of impaired cell growth isimportant in addressing the problem causing impaired growth. Forexample, early detection may allow for ambient conditions to be modifiedbefore cells experience irreversible damage. Early detection of aninfection affecting cells may allow for control measures to be taken toprevent spread of the infection to other cells or specimens.

Continuous monitoring of the rate of cell growth may also be used tosupport research related to the use of medications. When medications orother interventions such as genetic modifications intended orpotentially capable of cell growth or metabolism are investigated,continuous monitoring can quantify their effect on cells and provide ameasure of the kinetics of their action. Impairment in cell growth ormetabolic activity, or increased rate of cell growth may be adverseeffects of drugs. Continuous monitoring of cell growth may allow forbetter detecting such effects.

With the advent of technology, cells are often cultured in smallcompartments such as wells, which are more difficult to visuallyinspect. Spectrometry is used to measure cell concentration in a liquidmedium but this requires manually positioning of wells or othercontainers in a designated device which is not located within theincubator. This is labor intensive, increases the risk of mistakes andrequires displacement of cells from their preferred culture environment,which may have adverse effects on cell growth. Automated agar plateinoculation systems exist that perform seeding of microbiologic sampleson plates and monitor their growth using a camera but these do not useinfrared cameras, and are not commonly used in the research setting dueto their inflexibility and high price.

SUMMARY

In one embodiment, the present invention is directed to an apparatus formonitoring a cell culture comprising a) one or more infrared sensorspositioned adjacent to a cell culture, the one or more infrared sensorscapable of recording an infrared heat signal from the cell culture; b) apower source in electrical communication with the one or more infraredsensors; c) a data storage and computation device configured to receiveand analyze the infrared heat signal from the one or more infraredsensors; and d) a transmitter device in electrical communication withthe storage and computation device; wherein the apparatus monitors thepattern of heat production in the cell culture.

In another embodiment, the present invention is directed to a method formonitoring and analyzing the metabolic activity of cells comprising thesteps of: a) recording high definition images of infrared spectrumemission from the cells and from a reference sample using theabove-described apparatus; b) analyzing the images by computerized imagerecognition to determine at least the amount of heat produced by thecells, the regions of the culture where the heat is produced, and thechange in heat production over time to provide an uncorrected analysis;and c) optionally correcting the analysis by comparing the images of thecells with the images of the reference sample to provide a correctedanalysis.

These and other embodiments will become apparent from the followingdetailed description of the invention.

BRIEF DESCRIPTION OF THE DRAWINGS

The subject matter which is regarded as the invention is particularlypointed out and distinctly claimed in the claims at the conclusion ofthe specification. The foregoing and other features, and advantages ofthe invention are apparent from the following detailed description takenin conjunction with the accompanying drawings in which:

FIG. 1 illustrates one embodiment of the cell growth monitoring systemof the invention;

FIG. 2 illustrates another embodiment of the cell growth monitoringsystem of the invention including an amplifier and a control unit;

FIG. 3 illustrates another embodiment of the cell growth monitoringsystem of the invention including a reference component;

FIG. 4 illustrates another embodiment of a cell growth monitoring of theinvention including a sensor array; and

FIG. 5 illustrates an infrared system for cell culture monitoringincluding an infrared lens in accordance with the present invention.

DETAILED DESCRIPTION

There is a need for automated, low cost means to monitor cell growth tosave time, reduce labor associated with lab work, save money lost due towaste of reagents and equipment, and shorten time to microbiologicresults that may affect patient treatment and outcomes.

The present invention relates to an apparatus and method for infraredmonitoring of cell cultures, and more specifically to an apparatus andmethod that utilizes infrared sensors or cameras to detect changes inthe pattern of heat production within metabolically active cells. Thepresent invention offers the advantages of automation, ease of use, lowcost, and speed of data acquisition that could improve cell culturegrowth conditions, patient treatment, and outcomes.

As indicated above and shown in FIG. 1, the present invention isdirected to an apparatus for monitoring the pattern of heat productionin a cell culture. The apparatus comprises (a) one or more infraredsensors 101 positioned adjacent to a cell culture 102; (b) a powersource 103 in electrical communication with the infrared sensors 101;(c) a data storage and computation device 104 configured to receive andanalyze the infrared heat signal from the sensors; and (d) a transmitterdevice 105 in electrical communication with the storage and computationdevice 104. Each of these components is described in more detail below.

The apparatus of the present invention includes one or more infrared(IR) sensors. The infrared sensors used in the apparatus of theinvention may be any sensor that effectively senses infrared radiation(e.g. radiation from 700 nm-1 mm). Examples of useful sensors that maybe implemented in the apparatus of the invention include, but are notlimited to, cameras, bolometers, semiconductors, and the like. In onepreferred embodiment, the IR sensor is a camera, such as a camera in theconventional sense, with a transparent viewing port, and can incorporatelenses or other means for focusing incoming radiation. In oneembodiment, the IR sensors can be cameras which record high definitionimages, wherein high definition can be defined as being on the order of80×60 pixels or more. The IR sensors can include bolometers, whichmeasure the power of incident radiation by the measuring the heating ofa material with temperature-dependent electrical properties. The IRsensors can include semiconductors sensitive to IR light. The IR sensorscan be sensitive to many wavelengths of IR radiation, such as far IR(e.g. radiation from 15000 nm-1 mm), near IR (e.g. radiation from700-3000 nm), or any wavelength band in between.

The IR sensors of the invention are preferably passive IR sensors,defined as sensors that measure IR light radiating from objects, whereinthe sensor itself does not radiate energy at the sample for detectionpurposes. A passive IR sensor detects changes in the amount of IRradiation impinging upon it, which varies depending on the temperatureand surface characteristics of the objects in front of the sensor. Thesensor converts the resulting change in incoming IR radiation into achange in output value which can be used to detect and record the IRradiation.

The cell culture can be contained in a culture vessel. In oneembodiment, the culture vessel can be a container such as a petri dish,test tube, flask, and the like. In other embodiments, the culture vesselcan be a well, which can be part of a multi well plate, or can be thewell plate itself. The culture vessel can be sealed, closed with a lid,or open to the environment on at least one surface. In some embodiments,the culture vessel is open on the top, such as an open well with no lid.In other embodiments, the culture vessel is closed but has one or moreaccess ports allowing inflow or outflow of material, including liquids,solids, gases, suspensions, and the like.

The IR sensors can be placed adjacent to the sample which can be cellculture, medium, or reference sample, and which can be contained in aculture vessel. As defined herein, “adjacent” means any orientation thatis sufficiently close and properly situated to detect IR radiationemitted from the sample. For example, the sensor can be placed above,underneath, or on any side of the sample. In one embodiment, the sensoris placed above the sample, wherein the sample is a cell culture. Inanother embodiment, the sensor can be placed directly on the outersurface of a culture vessel containing the sample. In anotherembodiment, the sensor is placed a short distance away from the sample,for example less than 100 cm, less than 50 cm, or less than 30 cm. Insome embodiments, the IR sensor can be placed on the lid of the culturevessel, such as a petri dish, or replace the lid of the culture vessel.In some embodiments, the sensor can be placed inside the culture vessel,and is optionally immersed in the medium containing the sample.

IR radiation from the culture vessel can be detected by a single IRsensor, or by an array of IR sensors, for example 2, 3, 4, 5, 6, or moreIR sensors. The array of IR sensors or a single IR sensor can bearranged to detect IR radiation from different regions of the cellculture. The culture vessel must have a structure, or at least a portionof its structure, that is transparent to IR radiation so that the IRsensors can detect IR emissions through the wall of the culture vessel,unless the IR sensors are located inside the culture vessel.“Transparent to IR” is defined here as a material that permits nearly100%, at least 95% or at least 90% transmission of infrared radiation.For example, the culture vessel may have at least a portion of itsstructure made of IR transparent material such as chalcogenide glass, IRgrade fused silica, sapphire, calcium fluoride, silicone, zinc sulphite,cadmium telluride, quartz, IR transparent plastics, and the like.

The apparatus according to the present invention includes a power sourcein electrical communication with the IR sensors. Power sources suitablefor implementation in the apparatus and methods of the invention can beof a kind that store power, such as batteries or capacitors, or receivepower without wires, such as by solar power or inductive coupling. Insome embodiments the power source receives power through a wiredconnection with an electrical main, external battery, externalgenerator, or the like. In some embodiments, the power source can be inelectrical communication with an amplifier, a data storage andcomputation device, a transmitter, or a control unit. In someembodiments, a single power source supplies power to all the componentsof the apparatus which require power. In other embodiments, there can bemultiple power sources, such that some power sources supply somecomponents and other power sources supply other components.

A data storage and computation device is in communication with thesensor and/or amplifier, and is configured to receive and analyze the IRsignal. The data storage and computation device can be a standarddigital computer, or simply a printed circuit board and memory device,or other devices that accomplish the same function. The data storage andcomputation device can also be in communication with a power supply, atransmitter, and a control unit. The data storage and computation devicecan analyze the data in the IR signal to generate an analyzed signal orfurther analyze to produce image data which can be viewed or sent toother devices.

In some embodiments a transmitter can be in communication with thesensor, amplifier, data storage and communication device, control unit,and/or power source. As shown in FIG. 2, in one embodiment, thetransmitter 105 receives information from the data storage andcomputation device 104 and transmits the information to a control unit107. The control unit 107 can exchange information from the computationdevice 104 via wired connection e.g. ethernet, or alternately throughthe transmitter-receiver 105. The transmitter can send the informationvia a wired connection or by wireless transmission. The wirelesstransmission can be by technology such as BLUETOOTH (short link radiotechnology), WiFi, RFID, or the like.

As shown in FIG. 2, the apparatus includes an amplifier 106 incommunication with the IR sensor 101. The amplifier 106 can amplify thesignal received by the IR sensor 101. The amplifier 106 can beincorporated with the sensor component or external to the sensor 101. Insome embodiments, the amplifier 106 is positioned between the IR sensor101 and the data storage and computation device 104.

As shown in FIG. 2, a control unit 107 receives information from othercomponents of the system. The control unit 107 can be any computationdevice known in the art such as a smartphone, laptop, or cloud service.The control unit 107 can receive IR signal information, and can analyzeand process it to produce a determination of heat production in the cellculture. In some embodiments, the control unit 107 is in communicationwith the data storage and computation device 104. In some embodiments,the control unit 107 produces an analysis of metabolic activity of thecell culture over time. In some embodiments, the functions oftransmitter 105 and data storage and computation device 104 areincorporated in the control unit 107; the control unit 107 in such anembodiment receives data from the IR sensors 101, analyzes it, and thenprocesses it to produce a determination of heat production in the cellculture.

In some embodiments of the apparatus of the invention, the control unitcan receive information other than IR heat signal. Such as informationcan include pH, humidity, and carbon dioxide concentration, or otherparameters related to cellular growth. This information can come fromsensors in addition to the IR sensors, for example pH sensors to detectpH.

In some embodiments the control unit can modify the growth conditions ofthe cell culture as desired by the user. For example, if the controlunit finds the growth of the cells is too slow, the control unit canadjust the temperature of the cell culture, and if the pH of the cellculture is out of the desired range, the control unit can adjust the CO₂concentration.

In some applications of the apparatus and method of the invention, itmay be desirable to compare the infrared signal from the cell culture toa reference sample. As shown in FIG. 3, the apparatus of the presentinvention may therefore also include embodiments which include areference component 200 with one or more reference samples 202. Inaddition to one or more reference samples, the reference component is incommunication with the control unit, includes one or more IR sensors,and can include an amplifier 206, transmitter, digital computer, andpower source configured similarly to the components used to monitor thecell culture. In one embodiment the system comprises one or morereference infrared sensors 201 positioned adjacent to a reference sample202; a reference power source 203 in electrical communication with thereference infrared sensors 201; a reference data storage and computationdevice 204 configured to receive and analyze the infrared heat signalfrom the reference infrared sensors 201; a reference transmitter device205 in electrical communication with the reference storage andcomputation device 204 and configured to transmit information to acontrol unit 107.

The reference samples can contain only media in a culture vessel, or cancontain both cells and media. The reference component allows comparisonof the pattern of heat production in the cell culture with the patternof heat production in the reference sample. This comparison can be usedto reduce signal noise or to correct the IR signal from thenon-reference cell culture. In some embodiments, the reference sampleincludes cells different from the cell culture sample and is used tocompare the heat production of different cell types. In otherembodiments the reference sample contains cells similar to those in thecell culture, but grown under different conditions.

As shown in FIG. 4, in some embodiments of the invention one or more IRsensors may comprise a sensor array 301. In this embodiment, the sensorarray 301 comprises multiple individual IR sensors 302 which are used tomonitor heat production caused by metabolism and growth of a cellculture 303 in a petri dish 304.

Sensor array 301 can comprise 2, 3, 4, 5, 6, or more individual sensors302. The sensors in the sensor array can be arranged in any suitablepattern which allows measurement of the desired regions of the cellculture. In some embodiments, parts of the sensor array measure IRsignal from the portion of the cell culture with cells, and portions ofthe sensor array measure IR signal from a portion of the cell culturewithout cells.

As shown in FIG. 5, in one preferred embodiment of the invention aninfrared camera is used to monitor the growth of a cell culture. A cellculture 403 creates heat as it grows on a growth medium 404 in a culturedish 405, wherein culture dish 405 is covered on its top surface by aninfrared transparent lid 402. The creation of heat by cell culture 403results in infrared emission from the cell culture that is greater thanthe surrounding culture dish 405, medium 404 and lid 402. An infraredcamera 400 is positioned adjacent to lid 402, with a lens 401 interposedbetween camera 400 and lid 402. Infrared camera 400 is in electricalcommunication with computer 406 which has a network connection 407. Inone embodiment, one or more lenses 401 are positioned between the IRcamera 400 and cell culture 403.

An infrared transparent lid 402 is preferably chosen with low emissivityto allow the infrared emission from the culture to be visible to theinfrared camera 400. In some embodiments the entire lid 402 is IRtransparent. In other embodiments, a portion of the lid 402 is IRtransparent to form an IR window, and the IR camera 400 is situated todetect IR emission that passes from the cell culture 403 through the IRwindow. The lens 401 is preferably chosen so that it is transparent toand can focus IR radiation. Such a lens may be termed an IR lens. In oneembodiment the lid of the culture vessel comprises an IR transparentmaterial configured to function as a lens.

In some embodiments of the invention the cell culture is grown in anenclosure such as an incubator. In such embodiments, all the componentsof the apparatus can be entirely outside the incubator (for example, theIR sensor can detect IR from the cell culture through a window or otherIR transparent surface of the incubator), or some components can beinside the incubator and some outside. For example, in an embodiment thecomputer, control unit, and power source can be outside the incubatorand the transmitter, IR sensor, and amplifier are inside the incubator.

The cell culture that may be monitored using the apparatus and method ofthe invention may be one or more cells in a growth medium, the cells ofa kind that emits IR radiation when metabolically active. The cells canbe eukaryotic, prokaryotic, or archaea. If eukaryotic, the cells can befrom a one celled or multicellular organism. If eukaryotic, the cellscan be connected to each other as part of a tissue, and can be acomplete organism including an embryo. In some embodiments, the cellscan be an embryo generated as part of an in vitro fertilization (IVF)procedure.

The cells can be grown in any medium known in the art, for example amedium which is solid or semisolid, such as agar. For a solid orsemisolid medium, the cells can be grown on the surface of the medium orin the interior of the medium. In some embodiments the cells can begrown in a liquid medium, such as a liquid nutrient broth.

In another embodiment of the invention, the apparatus may be implementedin a method for monitoring and analyzing the metabolic activity of cellscomprising the steps of: a) recording high definition images of infraredspectrum emission from the cells and from a reference sample using theapparatus of the invention; b) analyzing the images by computerizedimage recognition to determine at least the amount of heat produced bythe cells, the regions of the culture where the heat is produced, andthe change in heat production over time to provide an analysis; and c)optionally correcting the analysis by comparing the images of the cellswith the images of the reference sample to provide a corrected analysis.

In one embodiment of the method of the invention, the recording stepcomprises detecting the IR light with an IR sensor to produce a highdefinition IR image on the sensor, and converting the high definitioninfrared image on the sensor to a digital image that is relayed directlyor indirectly to a computing device, and may be printed, scanned, ordisplayed to produce a form of recording visible to a user.

In one embodiment of the method of the invention, analyzing the imagescomprises windowing (sub-section) of the image and interpreting theimage relative to previous images to detect whether culture growth isproceeding as intended. A variety of image processing methods can beused to evaluate the infrared images that will be recognized by thoseskilled in the art as well as automatically by image recognitiontechnology used by computerized systems trained for this purpose. Theraw infrared image is preferably first processed using signal gain andoffsets and lookup table balancing to give high contrast to the desiredculture features in the image. In some embodiments the images ofinfrared spectrum emission are coupled with other signals to interpretthe infrared emission pattern or to augment or verify interferences fromthe infrared pattern. The other signals can include signals such as gasanalysis, spectroscopy, colorimetry, visible or ultraviolet light, andthe like.

In some embodiments, the analyzing step can further comprise determiningthe approximate size of the culture by counting the number of now highcontrast elements in the image (blob finding). Culture sizes can becompared to results in previous images taken over time to determine therate of growth of the sample. In other embodiments, sequential filteredimages may be subtracted from previous images and the resultant pixelscounted to determine the differential growth. Those skilled in the artwill recognize that a wide variety of methods similar to those outlinedabove can be used to determine the change in culture size.

Analysis of IR images can allow for identification of locations in whichheat is produced by metabolically active cells and monitoring this heatproduction over time. In some embodiments this allows for the detectionof early stage bacterial growth, estimation of the inoculum, predictingthe type of microorganism(s) growing by the spatial pattern of growthand the growth rate, identification of metabolic responses tointerventions (such as exposing cells to a drug, change in nutrientsprovided or manipulating gene expression), as well as early detection ofmalfunction of cells due to a disease (e.g., viral infection of cells)or in plausible ambient conditions. The apparatus and method of thepresent invention can potentially add to the ability to differentiatehealthy from sick tissues sampled from a human or animal body. Thepresent invention can allow faster evaluation of cell growth than ispossible by simple visual inspection. Other methods of determining cellmetabolism and growth involve measurement of gas such as CO₂ produced byliving cells, but these methods would not work for unsealed containerssuch as petri dishes, whereas the apparatus and method of the presentinvention can work in a sealed or unsealed environment.

In some embodiments the apparatus is used to detect cell growth anddetermine the rate of cell growth or cell metabolism. In otherembodiments the apparatus is used to estimate the initial inoculum ormicroorganism taxa. In other embodiments the apparatus is used toevaluate cell response to a drug. Cell response to a drug can bemeasured by the implanting of drug-infused discs in the cell culture,and subsequent measurement of the reduction in rate or extent of growthor metabolism. In some embodiments the evaluation of cell response to adrug measures antibiotic susceptibility.

In another embodiment, the analyzing step comprises processing toincrease contrast between cells and medium to create higher contrastelements corresponding to the cells in the image; and then counting thehigher contrast elements in the image to provide an estimate of themetabolic activity of the cells; and then comparing metabolic activityover time to determine the rate of growth of the cells.

According to one embodiment of the invention, an apparatus formonitoring a cell culture comprises a) one or more infrared sensorspositioned adjacent to a cell culture, said one or more infrared sensorscapable of recording an infrared heat signal from said cell culture; b)a power source in electrical communication with said one or moreinfrared sensors; c) a data storage and computation device configured toreceive and analyze said infrared heat signal from said one or moreinfrared sensors; and d) a transmitter device in electricalcommunication with said storage and computation device; wherein saidapparatus monitors the pattern of heat production in said cell culture.

In another embodiment, said one or more infrared sensors comprise one ormore infrared cameras.

In another embodiment, the apparatus further comprises an amplifierpositioned between said one or more infrared sensors and said datastorage and computation device.

In another embodiment, the apparatus further comprises a control unit incommunication with said data storage and computation device.

In another embodiment, the apparatus further comprises a referencecomponent in communication with said control unit, said referencecomponent comprising a) one or more infrared sensors positioned adjacentto a reference sample, said one or more infrared sensors capable ofrecording an infrared heat signal from said reference sample; b) a powersource in electrical communication with said one or more infraredsensors; c) a data storage and computation device configured to receiveand analyze said infrared heat signal from said one or more infraredsensors; and d) a transmitter device in electrical communication withsaid storage and computation device; wherein said reference componentmonitors the pattern of heat production in said reference sample andwherein said control unit compares said pattern of heat production insaid cell culture with said pattern of heat production in said referencesample.

In another embodiment, said cell culture is contained in a culturevessel.

In another embodiment, said culture vessel is a petri dish.

In another embodiment, said culture vessel is a flask.

In another embodiment, said culture vessel is a test tube.

In another embodiment, said culture vessel is a well plate.

In another embodiment, said culture vessel comprises infraredtransparent materials.

In another embodiment, said one or more infrared sensors are positionedabove said cell culture.

In another embodiment, the apparatus further comprises one or morelenses positioned between said one or more infrared sensors and saidcell culture.

In another embodiment, the infrared sensors are located on the outersurface of the culture vessels.

In another embodiment, the infrared sensors are located on the lids orreplacing the lids of the culture vessels.

In another embodiment, the apparatus further comprises a component forwired or wireless data transmission to a remote computer, handhelddevice, or cloud.

In another embodiment, the infrared sensors can record continuously orintermittently.

In another embodiment, the infrared sensors can be sterilized usingtechniques that do not damage said sensors.

In another embodiment, the infrared sensors and cell cultures are bothlocated in a cell culture incubator.

In another embodiment, the control unit can receive information otherthan heat signal, wherein said information other than heat signal caninclude pH, humidity, and carbon dioxide concentration.

In another embodiment, the control unit can modify the growth conditionsof the cell culture.

In another embodiment, the images of infrared spectrum emission are usedto detect cell growth.

In another embodiment, the images of infrared spectrum emission are usedto monitor the effect of a drug.

In another embodiment, the images of infrared spectrum emission are usedto detect antibiotic susceptibility.

In another embodiment, the images of infrared spectrum emission are usedto estimate the initial inoculum or microorganism taxa.

In another embodiment, the images of infrared spectrum emission arecoupled with other signals to interpret the infrared emission pattern orto augment or verify interferences from the infrared pattern; whereinthe other signals can comprise gas analysis, spectroscopy, colorimetry,or visible or ultraviolet light.

In another embodiment, the cells are eukaryotic, prokaryotic, orarchaea.

The flowchart and block diagrams in the Figures illustrate thearchitecture, functionality, and operation of possible implementationsof systems, methods, and computer program products according to variousembodiments of the present invention. In this regard, each block in theflowchart or block diagrams may represent a module, segment, or portionof instructions, which comprises one or more executable instructions forimplementing the specified logical function(s). In some alternativeimplementations, the functions noted in the block may occur out of theorder noted in the figures. For example, two blocks shown in successionmay, in fact, be executed substantially concurrently, or the blocks maysometimes be executed in the reverse order, depending upon thefunctionality involved. It will also be noted that each block of theblock diagrams and/or flowchart illustration, and combinations of blocksin the block diagrams and/or flowchart illustration, can be implementedby special purpose hardware-based systems that perform the specifiedfunctions or acts or carry out combinations of special purpose hardwareand computer instructions.

The descriptions of the various embodiments of the present inventionhave been presented for purposes of illustration, but are not intendedto be exhaustive or limited to the embodiments disclosed. Manymodifications and variations will be apparent to those of ordinary skillin the art without departing from the scope and spirit of the describedembodiments. The terminology used herein was chosen to best explain theprinciples of the embodiments, the practical application or technicalimprovement over technologies found in the marketplace, or to enableothers of ordinary skill in the art to understand the embodimentsdisclosed herein.

What is claimed is:
 1. A method for monitoring and analyzing metabolicactivity of a cell culture, the method comprising the steps of:recording first high definition images of infrared spectrum emissionfrom the cell culture over time in a medium and second high definitionimages of infrared spectrum emission from a reference sample using highdefinition infrared sensors, the reference sample comprising a secondcell culture having at least one of a different cell type or a same celltype grown under different conditions, the cell culture in a culturevessel, the high definition infrared sensors comprising a lid of theculture vessel configured as an infrared lens; analyzing the first highdefinition images by computerized image recognition to determine atleast an amount of heat produced by the cell culture, two or moredifferent regions of the cell culture where heat is produced, and achange in heat production in the two or more different regions over timeto provide an analysis, wherein the two or more different regionsinclude a first region of the cell culture having cells and a secondregion of the cell culture without cells; correcting the analysis bycomparing the first high definition images of infrared spectrum emissionof the cells with the second high definition images of infrared spectrumemission of the reference sample to provide a corrected analysiscomprising reduced signal noise; providing a metabolism analysis basedupon the corrected analysis, wherein the metabolism analysis comprisesdetermining a change in metabolism based upon the change in heatproduction, wherein an increase in heat production is correlated with anincrease in metabolism and a decrease in heat production is correlatedwith a decrease in metabolism; processing the first high definitionimages to increase contrast between cells and the medium to createhigher contrast elements corresponding to the cells in the images;windowing a first image of the first high definition images; windowing asecond image of the first high definition images, wherein the secondimage is a previous image relative to the first image; counting thehigher contrast elements in the images to determine a size of the cellculture; comparing a number of high contrast elements in the first imageto a number of high contrast elements in the second image to determine arate of growth of the cell culture based on a change in culture sizeover time; and modifying a growth condition of the cells in response tothe rate of growth of the cell culture falling outside of a desiredrange, wherein modifying the growth condition comprises at least one ofadjusting a temperature of the cell culture and adjusting a CO₂concentration of the cell culture to increase or decrease the rate ofgrowth.
 2. The method of claim 1, comprising monitoring the effect of adrug.
 3. The method of claim 1, comprising detecting antibioticsusceptibility.
 4. The method of claim 1, comprising estimating theinitial inoculum or microorganism taxa.
 5. A method for monitoring andanalyzing metabolic activity of cells in a cell culture comprising thesteps of: recording first high definition images of infrared spectrumemission from the cell culture over time in a medium and second highdefinition images of infrared spectrum emission from a reference sampleusing high definition infrared sensors, the reference sample comprisinga second cell culture having at least one of a different cell type or asame cell type grown under different conditions the cell culture in aculture vessel, the high definition infrared sensors comprising a lid ofthe culture vessel configured as an infrared lens; analyzing the firsthigh definition images by computerized image recognition to determine atleast an amount of heat produced by the cell culture, two or moredifferent regions of the cell culture where heat is produced, and achange in heat production in the two or more different regions over timeto provide an analysis, wherein the two or more different regionsinclude a first region of the cell culture having cells and a secondregion of the cell culture without cells; providing a metabolismanalysis based upon the analysis, wherein the metabolism analysiscomprises a determination of a change in metabolism based upon thechange in heat production, wherein an increase in heat production iscorrelated with an increase in metabolism and a decrease in heatproduction is correlated with a decrease in metabolism processing thefirst high definition images to increase contrast between cells and themedium to create higher contrast elements corresponding to the cells inthe images; windowing a first image of the first high definition images;windowing a second image of the first high definition images, whereinthe second image is a previous image relative to the first imagecounting the higher contrast elements in the images to determine a sizeof the cell culture; comparing a number of high contrast elements in thefirst image to a number of high contrast elements in the second image todetermine a rate of growth of the cell culture based on a change inculture size over time; and modifying a growth condition of the cells inresponse to the rate of growth of the cell culture falling outside of adesired range, wherein modifying the growth condition comprises at leastone of adjusting a temperature of the cell culture and adjusting a CO₂concentration of the cell culture to increase or decrease the rate ofgrowth.
 6. The method of claim 5, further comprising coupling each ofthe first image and the second image with a signal selected from thegroup consisting of a gas analysis signal, a spectroscopy signal, acolorimetry signal, a visible light signal, and a ultraviolet lightsignal to interpret the infrared emission pattern or to augment orverify interferences from the infrared pattern.
 7. The method accordingto claim 5, wherein analyzing the first high definition images comprisesprocessing the first high definition images with signal gain, offsets,and look up table balancing to create high contrast image features.